Site-specific Antibody Service


  • PTMs (Post-translational Modifications) are an important consideration in research. They contribute to a diversity of biological activities and effect protein structure and cellular functions. Protein phosphorylation, methylation, and aceytylation are the most common types of modifications of the more than 200 different attached modifications known to exist in vivo.
  • These site-specific antibodies have a number of potential applications including:cell signaling, protein structure, function, development, maturation, localization, disease diagnostics, activity, and many more. ABclonal is a certifiable expert in the development of thousands of custom site-specific antibody services over the years.
  • All custom modification antibody production will undergo double affinity purification with modified and unmodified peptide.

PTMs in the Cell

The major types of post-translational modifications (Jensen O. N. 2006)

One-Stop Antibody Service


ABclonal Services Available


Phosphorylation Acetylation Methylation Ubiquitination
Citrullination Succinylation Alkylation Isoprenylation
Prenylation Sumoylation Tyrosine sulfation Tyrosine nitration
Glycosylation Biotinylation Polyglutamylation Other PTMs

Package


  • Peptide design and synthesis
  • Customer supplied peptide
  • Peptide-KLH conjugation
  • Immunization of 2 rabbits
  • ELISA titer test
  • Antigen affinity column purification

Note:

1.If the antigen is provided by the customer: modified peptide (>15 aa >90% purity, 15 mg) and unmodified control (>15 aa >90% purity, 11 mg).

2.The success rate of the modified antibody preparation is lower than the normal one. To ensure the probability of obtaining a useful antibody after immunization, we recommend to immune 3-4 rabbits according to our years of accumulated experience.

Modified peptide synthesis usually with a length of about 20 amino acids.Approximately 10mg are used in modified epitope affinity purification, 10mg used in antibody production, and 10mg used in QC.

Approximately 10mg non-modified peptide (about 20 amino acids) used in affinity purification.

To obtain a more specific modified antibody, sometimes we use shortened modified peptides to purify.

Professional antigen design(free)MORE


Proper design of a suitable antigen is the most critical step of getting an useful antibody.

High-quality antigen preparationMORE


We choose the best region to synthesis peptide coupling KLH as antigen to immune animals through professional analysis.

Each synthesized peptide goes through HLPC and MS tests to ultimately obtain a useful antibody.

Other Modifications


ABclonal offers DNA methylation, RNA methylation, pan-acetylation, non-histone modification and other custom specific modified antibodies that are developed to meet your research needs.

Use Nipponbare callus (same quantity, about 3g) as sample material of ChIP experiment, 02g and 03g as target gene. Conduct IP experiment using Anti- H3K27me3 (ABclonal) and Anti- H3K27me3 (Company M).

Conclusion:
The efficiency of abclonal anti- H3K27me3 immunoprecipitation is 10 times that of company M anti- H3K27me3.

High Throughput Detection


Obtaining a useful site-specific antibody is not easy and most manufacturer can not achieve because of the antibody validation. ABclonal has built advanced testing platforms for Western blot, immunohistochemistry, and immunofluorescence-based assays, supplemented by immunoprecipitation, RIP, ChIP ,ChIP- seq, and shRNA knock- out validation. In additional to manufacture, we dedicate to be the best validation platform in the world. We take every measure to provide the most accurate and reliable data for each of our customers.

ABclonal's standard option for polyclonal antibody is optimized to produce antiserum in the shortest possible time. It is the most affordable option, popular among researchers.

The protocol uses two New Zealand White rabbits that are Specific Pathogen-Free. Injections are subcutaneous (SQ) as emulsions in Complete Freund's Adjuvant (CFA) or Incomplete Freund's Adjuvant (IFA).We guarantee that a single antigen will be injected into both animals.

Procedure Protocol Day Description
Control Serum Collection Day 0 Pre-immune bleed (5mL per rabbit)
Primary Injection Day 1 Immunize with 0.50mg of antigen in CFA, 10 SQ sites
1st Booster Day 14 Boost with 0.25mg of antigen in IFA, 4 SQ sites
2nd Booster Day 28 Boost with 0.25mg of antigen in IFA, 4 SQ sites
Serum Collection Day 35 Bleed (~25mL per rabbit)
3rd Booster Day 42 Boost with 0.25mg of antigen in IFA, 4 SQ sites
Serum Collection Day 56, 58 Two bleeds (~50mL total per rabbit)
ELISA Test Day 60 ELISA titration (results available online);
Verify disposition of rabbits; decide to continue or terminate
Purification Optional 6-10 mg purified antibodies
Note: After Day 35, the Scheduled days in this protocol are approximate. They depend on the titer test.
Quote
  • For quotation requests and questions, you may contact us by email, phone, or via our online quotation system.
  • Orders can be placed by phone, email, fax, or online with a formal Purchase Order or credit card.
  • Please send your samples to ABclonal, 86 Cummings Park, Woburn, Massachusetts.
  • Our customer service is available 24hours per day/7 days a week/365 days a year.

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